Recombinant human cc10 protein for treating influenza

FIELD: medicine.

SUBSTANCE: invention refers to medicine and concerns using rhCC10 protein for preparing a therapeutic agent for the therapeutic or preventive effect on influenza virus.

EFFECT: invention provides reducing a pulmonary titre of influenza virus.

11 cl, 4 dwg, 1 tbl, 5 ex

 

The technical field to which the invention relates

Embodiments of the present invention relate to methods of reducing virus titers in vivo and treatment of patients with viral respiratory infections. Embodiments of the present invention also include methods of treating influenza infection, including influenza, especially H1N1. In addition, embodiments of the present invention relate to methods for treating the above infections with the use of injected intranasally, and/or intravenous and/or entered by the inhalation of recombinant human protein S.

The level of technology

Protein of Clara cells with a molecular weight of 10 kDa (SS) or uteroglobin (UG) is a small homodimeric secretory protein produced by certain epithelia of the mucous membranes and other organs of epithelial origin (Mukherjee, 1999). Protein S consists of containing 70 amino acid residues of two identical subunits, each of which is the leitmotif of the secondary structure forms a "chetyrehspalnyh node, connected in anti-parallel orientation of the two disulfide bonds between cysteines 3 and 69', 3' and 69 (Matthews, 1994; Morize, 1997). Protein S - the first representative of a growing family of small globular proteins with the same secondary, tertiary and even�artichow patterns and, believed to perform similar functions. It turns out that his primary form is C) an homodimer, containing two disulfide bonds. In humans, the main place of production of the protein S are easy, although some of the other bodies are synthesized small amounts of mRNA that encodes this protein (Singh, 1987; Sandmoller, 1994). Protein S is an anti-inflammatory and immunomodulatory protein that has been characterized by its various interactions with other proteins, receptors and cell types (see review in Mukherjee, 1999, and Pilon, 2000). The decreased levels of the protein products SS or mRNA were detected in different tissue samples and biological fluids in a wide range of disease States characterized by some degree of inflammation, including pneumonia (Nomori, 1995).

Physiology of protein S in different types of pulmonary infection was investigated in one line of mice with a knockout of protein S. In two studies, mice with a knockout of protein S and mouse wild-type were infected with two typical human respiratory pathogens - Pseudomonas aeruginosa, or adenovirus. The knockout mice were found more rapid disappearance of pathogens with greater neutralization of pathogens is the innate immune system, which suggested the use of protein deficiency S during viral and bacterial �Peccei (Hayashida, 1999; Harrod, 1998). This is consistent with earlier observations, in which it was observed that the protein S is immunosuppressive agent (Dierynck, 1995; 1996). This indicates that the protein S able to suppress the natural immune response to infection, bacterial or viral, including influenza. Therefore, you should not expect that the introduction of the protein S in the presence of viral or bacterial respiratory infection benefit the patient. Subsequently, it was noted that recovery of function S in knockout mice using recombinant human protein S (rhCC10), inserted before infection with respiratory syncytial virus (RSV), leads to faster disappearance of the infection than in knockout mice without such reduction (Wang, 2003). However, recent studies have shown that rhCC10 protein can prevent the development of acquired immunity, particularly the emergence of antigen-specific T cells, when present simultaneously with the interaction of antigen with dendritic cells (Johansson, 2007). This again indicates that the introduction of rhCC10 protein may not be useful for a patient with an infection. Thus, in present knowledge concerning potential hazards or use applications rhCC10 protein for the treatment of respiratory infections, contradictory and do not allow to draw conclusions �otnositelno safety and/or effectiveness of the use of protein S to treat various types of respiratory infections. There is no information concerning the action of protein S for influenza infections. In this application is described: a direct test of the effectiveness of rhCC10 protein of influenza type A in vivo, a direct test of antiviral effect of protein S at the cellular level, the mechanism of its action and its possible application for the treatment and/or prophylaxis of viral infections, in particular influenza infection.

The flu virus over the last 120 years was the cause of the four major outbreaks (1889, 1918, 1957 and 1968), led to the deaths worldwide, about 50÷100 million people. The flu virus is orthomyxovirus, RNA-containing virus that is transferred by aerosol and by direct contact of infected surfaces to the mucosa of the nose and infects epithelial cells of the respiratory tract. Influenza infection can cause severe symptoms, including fever, sore throat, muscle pain, malaise, weight loss, respiratory congestion, and in some cases respiratory failure and death. The flu causes the acquired immune response (cytotoxic T-cells and antibodies), which is usually the normal healthy individuals eliminates infection within 1÷2 weeks. Some subtypes of influenza infecting humans, including avian flu (H5N1), seasonal influenza (H3N2) and swine flu (H1N1) may locationcalifornia means, such as neuraminidase inhibitors. However, the high mutation rate of influenza virus has led to the emergence of strains of drug-resistant (Moscona, 2009), so the widespread use of antiviral drugs for the prevention and/or treatment will accelerate the development of resistance to these drugs. Therefore, for the treatment, cure and prevention of influenza infection is a need for new therapeutic agents. Also, there are no approved therapies for the vast majority of viral infections of the respiratory tract or other body systems.

The objective of the invention

The following is a non-exhaustive list of goals achieved in embodiments of the present invention.

The purpose of embodiments of this invention is the reduction of pulmonary virus titer and through this treatment, cure or prevention of influenza infection, particularly influenza infection type a and in particular infection by the H1N1 strain of influenza virus.

A further objective of embodiments of this invention is the reduction of pulmonary virus titer and treatment, cure or prevention of influenza infection by introducing protein S by intravenous, inhaled or intranasal route or combination of routes of administration.

Another aim of embodiments of the Fig�termination is the reduction in virus titer and treatment, the cure or prevention of viral infection by introducing protein S by intravenous, inhaled or intranasal route, by oral, intravaginal or by a combination of routes of administration.

Another objective of embodiments of this invention is the suppression of virus replication at the cellular level through the use of protein S or other members of the family of secretagogues.

Disclosure of the invention

These and other objectives, features and advantages are achieved in embodiments of the present invention by introducing rhCC10 protein in the range specified dosages with appropriate intervals or in one dose to reduce the titer of the virus and the treatment, cure or prevention of a viral infection.

These and other objectives, features and advantages are achieved in embodiments of the present invention by introducing a protein S in the range specified dosages with appropriate intervals or in one dose, if a patient is diagnosed with a viral infection the symptoms characteristic of a particular virus, and/or by detection of virus by standard methods in patient samples by cultivation of the virus, immunological detection of virus and/or detection of viral nucleic acid.

These and other objectives, features and advantages are achieved also in parentadolescent of this invention by introducing protein S in the range specified dosages with appropriate intervals or in one dose, if a patient is diagnosed with influenza infection by symptoms of fever, myalgia and flushing and/or for detection of influenza virus by standard methods in patient samples (nasal swabs, blood samples or sputum) by culturing of the virus, immunological detection of virus and/or detection of viral nucleic acid.

In certain aspects of the invention protein S administered intranasally in a dose of approximately equally distributed between both nostrils, in the range from 1.5 μg to 1.5 mg per kilogram of body weight per day, or multiple doses, which together are in the range of daily dosages, to reduce pulmonary virus titer and treatment, cure or prevention of influenza infection.

In another aspect, the protein S administered intravenously at a dose of 10 mg per kilogram of body weight per day or in multiple doses, the total of which are in the range of daily dosages, for the treatment, cure or prevention of influenza infection.

In another aspect, the protein S superhuman origin is introduced in this range of dosages with appropriate intervals or in one dose, if a patient is diagnosed with a viral infection the symptoms characteristic of a particular virus, and/or by detection of the virus by standard methods in patient samples through cultivation of the virus, immuno�oricheskogo virus detection and/or detection of viral nucleic acid.

In yet another aspect, another representative of the family of proteins S, which are generally known as secretogranin, is introduced in this range of dosages with appropriate intervals or in one dose, if a patient is diagnosed with a viral infection the symptoms characteristic of a particular virus, and/or by detection of the virus by standard methods in patient samples through cultivation of the virus, immunological detection of virus and/or detection of viral nucleic acid.

Brief description of illustrations

Fig.1 - histogram, which presents the H1N1 viral load for the second day in the lungs of infected cotton hamsters (cotton rats), which was administered intranasally rhCC10 protein. The viral titer is expressed in units of (×107) TCID50per gram of tissue.

Fig.2 is the histogram, which presents the H1N1 viral load for the second day in the lungs of cotton hamsters that were injected intraperitoneally rhCC10 protein. The viral titer is expressed in units of (×107) TCID50per gram of tissue.

Fig.3 is the histogram, which shows the suppression of rhCC10 protein of virus replication in cell cultures. RhCC10 protein was added to the culture medium of cells Ner at concentrations of 100 μg/ml, 300 μg/ml and 1 mg/ml and left for 4 hours. Then, the medium was removed and replaced with fresh cells infected with RSV for 1 hour. Then the cells were washed for UDA�termination of excess virus, again added rhCC10 protein and incubated for another hour. Then cells were washed to remove excess protein SS. The virus titers in culture medium was measured on the fourth day after infection. Each protein concentration SS applied in three replicates.

Fig.4 - comparison of the histogram of the antiviral action of rhCC10 protein when administered before infection and after infection. Cells Nor treated with rhCC10 protein at a concentration of 1 mg/ml and infected with RSV, as in the case of Fig.3. In addition, 1 mg/ml rhCC10 protein was injected 1 hour after infection (treatment day 0), after 24 hours after infection (treatment day 1) and 48 hours after infection (treatment on day 2). The virus titers in culture medium was measured on the fourth day after infection.

The implementation of the invention

Embodiments of the present invention relate to the use of protein S to reduce pulmonary virus titer and to treat, cure or prevent influenza infection. Protein S preferably is a recombinant human protein S (rhCC10), obtained using the processes described in the publication No. 20030207795 patent applications in the USA and in the application PCT/US 09/43613, each of which is fully incorporated into the present application by reference, or by using any other process that allows to obtain rhCC10 protein pharmacopoeias�Oh qualification. RhCC10 protein in embodiments of the present invention can be entered together with, without, before or after other intranasal, pulmonary or systemic treatment.

Without limiting the possible processes of synthesis that can be used to produce human protein S, recombinant human protein S (also known as uteroglobin), active in suppressing viral replication in vitro and in vivo, synthesized and characterized as described in the publication No. 20030207795 patent application USA, which issued U.S. patent No. 7122344.

Drugs rhCC10 protein for intranasal administration as described in PCT/US 09/43613 represent further embodiments of the present invention that can be used to suppress viral replication in vivo, especially in the nasal passageways and cavities.

Dosage

In the treatment of influenza infection is preferably administered rhCC10 protein intranasally in each nostril 1 to 3 times a day for 7÷14 days, every other day for another 14 days, after which - if necessary. More preferably rhCC10 protein immediately after the patient has fever, myalgia and flushing or diagnosing influenza.

The manufacturing process of rhCC10 protein may contain the following stages: a) creation of a bacterial expression system capable expressive�ü rhCC10 protein; (b) inoculation of the seed fermenter containing a bacterial expression system for obtaining a fermentation culture; C) adding to the fermentation culture of the inducing agent to induce expression of rhCC10 protein in a bacterial expression system; d) retrieving expressed at the stage (C) rhCC10 protein; and (e) purification cured at stage (d) rhCC10 protein, where the purification step includes using at least one filter and at least one ion-exchange column, as described in the publication No. 20030207795 patent application USA. RhCC10 protein can also be expressed in other expressing system - bacterial, fungal, insect, mammalian, plant, and purified so that its characteristics meet the requirements of pharmaceutical product suitable for administration to humans using standard methods.

Characteristics and test results for rhCC10 protein pharmacopoeial qualification, according to the publication No. 20030207795 patent application USA, which can be used to reduce the titers of the virus include the following.

TestFeature
ColorTransparent, colorless
AppearanceNo turbidity
HomogeneityHomogeneous
Cleanliness>95%
The degree of aggregation<5%
SterilitySterile
Biological activityPositive
Bacterial nucleic acid<100 PG per dose
Mass spectroscopyOBS. 16110
pH5-8
Isoelectric focusing4,7±1
Free thiols<10% by mass
LAL-test<5 U/mg
Copper<16 µm

In another embodiment, rhCC10 protein according to the present invention, which suppresses viral replication and inhibits the enzyme phospholipase a2(PLA2as described in PU�likely No. 20030207795 patent application of the United States.

To achieve the desired results, further described below, made reference to the methods of administration described in the following embodiments of the invention.

In one of the embodiments of the invention can be entered intranasal dose or multiple doses of rhCC10 protein, equivalent to a dose in the range of from about 1.5 μg to about 5 mg per kilogram of body weight per day. In another embodiment, rhCC10 protein can be administered in a specified range of daily doses. In yet another embodiment, rhCC10 protein can be administered in a specified range of daily doses for at least 7 consecutive days. In another embodiment, rhCC10 protein can be administered in a specified range of daily doses for at least 14 consecutive days. In another embodiment, rhCC10 protein can be entered in the specified range of doses every other day for 30 consecutive days. In another embodiment, rhCC10 protein can be administered in decreasing doses daily for 10 consecutive days, and decreasing these doses are maximum dose of each introduction during the first 3 days, intermediate dose each introduction in the next 3 days and low dose each introduction in the last 4 days. In one embodiment, is�the implementation of the invention rhCC10 protein can be entered in the specified range of doses or decreasing doses up to three times daily, approximately every 8 hours.

In another embodiment, the dose of rhCC10 protein can be administered to the patient intranasally in the form of an aerosol, in the form of nasal spray or wash, or by applying a gel or cream, or otherwise drip into the nasal passages.

In another embodiment, the dose of rhCC10 protein can be administered to the patient inalatore in the form of an aerosol, spray or inhaler dosing, or otherwise direct introduction into the lungs and Airways.

In another embodiment of the invention in the treatment or prophylaxis of influenza infection protein rhCC10 is administered intravenously in doses from 15 mg to 20 mg per kilogram of body weight, 1 to 3 times daily, for 7÷14 days and then every other day for another 14 days, after which - if necessary. In another embodiment, rhCC10 protein you can enter decreasing doses daily for 10 consecutive days, and decreasing these doses are maximum dose of each introduction during the first 3 days, intermediate dose each introduction in the next 3 days and low dose each introduction in the last 4 days. In another embodiment, rhCC10 protein can be entered in the specified range of doses or decreasing doses up to three times in den�, approximately every 8 hours.

In another embodiment, the above doses of rhCC10 protein can be administered to the patient using a combination of intranasal, inhaled, and intravenous routes. In another embodiment, rhCC10 protein can be administered, in accordance with the methods described above, before, simultaneously or after antiviral therapy, antibiotic treatment, treatment decongestants, antihistamines, mucolytic, expectorant, suppressant mucus formation, a surfactant, a bronchodilator agent, vasoconstrictor, sedative cure for cavities or other standard treatment. In yet another embodiment, rhCC10 protein can be administered, in accordance with the methods described above, to reduce pulmonary virus titer and treatment, cure or prevention of influenza infection.

Doses of rhCC10 protein described above and methods of administration can be used daily, more than once a day, three times a day, every other day, or decrease the dose, depending on the gravity of the subject to treatment of influenza infection, the General health status of the patient and the presence of associated disease States. For example, the heavier the infection, the greater the amount of protein rhCC10 may be required for its effective treatment. Evident�, what the doctor is able to determine the appropriate dosages, formulations and routes of administration on the basis of symptoms and response to treatment of the patient within the parameters and dose ranges described in embodiments of the present invention.

Recipes

Intranasal formulations, devices and methods that rhCC10 protein you can enter the route described in the application PCT/US 09/43613, which is fully incorporated in this application by reference. Formulation of rhCC10 protein for intravenous administration is a solution with a concentration of 5.5 mg/ml in 0.9% physiological saline solution and is described in the publication No. 20030207795 patent application USA, which is fully incorporated in this application by reference.

Example 1

Reproduction and determining the titer of influenza virus

Prepare strain A/PR/8/34 influenza virus A (H1N1), purchased from American Bank of type culture (American Type Culture Collection, Manasass, Virginia, USA). The influenza virus propagated in MDCK cell cultures (number CCL-34 catalog of ADS) by infecting a monolayer of cells at the stage of 60% of the merger (in vials with a volume of 150 cm2) influenza virus at a multiplicity of infection (MOI) of 0.01. After 3 or 4 days, when developed cytopathic process and most of the cells separated from the surface of the vessel, remove cells and the supernatant. The cells removed by centrifugation (800 g), nodosa�full-time the liquid is filtered (pore size 0.45 μm) and centrifuged (18000 g) for 2 hours at 4°C to precipitate the virus. Precipitate the virus was resuspended in DMEM, the suspension was divided into aliquots and stored at -150°C. the Titer of influenza virus is determined by placing 0.1 ml of serial dilutions of viral samples in monolayers of MDCK cells in a plate of 96 cells cultured in the presence of 0.1% bovine serum albumin (BSA) and trypsin. After 3 days register cytopathic effect and is determined by the method of Kärber dose 50% infection in tissue culture (TCID50).

Processing of lung tissue for analysis of viral load

Aseptically remove sections of the left and right lobes of the lungs of infected mice hamsters and cotton, weighed and homogenized in 1 ml DMEM for 45 s, using the machine to destroy tissue (Biopspec Products Inc., model 985-370) in mode 5. The homogenates was centrifuged at 3000 g for 20 min, the Clarified supernatant is collected and stored at -150°C until use.

Determination of virus titers

The titers of the virus stocks of the virus and multiplied in homogeneo lungs determined by serial dilution followed by analysis of blackarachnia (PFA) or analysis of foci formation (FFA). The number blakebrough units (the FIGHT) and forming foci units (OUE) in 1 ml of the original sample is calculated before the start of the study. Retain one set of samples of the virus sufficient for the PFA and FFA, and after�stop studies determine the number of FIGHTS and AOE. Serial dilutions of cultured virus in clarified medium (DMEM with 1% BSA) were prepared in the range of dilutions from 101up to 108. Each dilution characterize using PFA and FFA. For cultured influenza virus usually get credits 107-109The FIGHT/ml.

Example 2

Intranasal introduction rhCC10 protein to reduce pulmonary virus titer of influenza

Cotton hamsters (S. hispidus), a kind vole is an animal model in which the influenza virus replicates and causes a mild respiratory infection (Ottolini, 2005). Animals infect by intranasal inoculation of influenza virus and pulmonary virus titers reached a maximum after 2 days (about 48 hours) after inoculation. This model is used to identify compounds that inhibit replication of influenza virus in vivo.

Not containing pathogens of cotton hamsters were obtained from Virion Systems, Inc. (Rockville, MD). Only 18 cotton hamsters (S. hispidus, age 6-8 weeks) were infected with influenza virus type A (A/PR/8/34), the H1N1 strain by intranasal inoculation, the dose for each animal was 107TCID50in a volume of 0.1 ml. of Six animals were given a placebo (0.9% NaCl), six animals were injected with 0.5 mg/kg rhCC10 protein and six animals were injected with 5.0 mg/CT rhCC10 protein by intranasal installation 2 hours before inoculation of the virus. Animals were scored on the second day after infection,�GDSs titers of virus are usually the most high, and in lung tissue was determined by viral load. Fig.1 shows the reduction in virus titer in the lung tissue that was observed in both groups that received a dose of rhCC10 protein. The titer of virus in the lungs is expressed in units of (×107) TCID50/gram of tissue.

Example 3

Systemic administration of rhCC10 protein to reduce pulmonary virus titer of influenza

Only 18 cotton hamsters (S. hispidus, age 6-8 weeks) were infected with influenza virus type A (A/PR/8/34), the H1N1 strain by intranasal inoculation, the dose for each animal was 107TCID50in a volume of 0.1 ml. of Six animals received placebo (physiological saline solution), six animals received 0.5 mg/kg rhCC10 protein and six animals received 5.0 mg/kg rhCC10 protein by intraperitoneal injection. Intraperitoneal route of administration provides a significant amount of circulating rhCC10 protein and mimics intravenous people. Each animal received a total of 6 doses of either placebo or rhCC10 protein approximately every 12 hours, including two doses (morning and evening) the day before infection, two doses per day of infection and 2 doses the next day after infection (3 doses before infection and 3 doses after infection). Animals were scored on the second day after infection, when virus titers are usually the most high, and in the lung tissue were determined by viral load. Fig.2 illustrates with�tistichesky a significant reduction (p< 0.01) and the titer of virus in the lung tissue, found in the group with the dose of 5 mg/kg rhCC10 protein, and a trend of decrease in viral titer in the group with a dose of 0.5 mg/kg. virus Titer in the lungs is expressed in units of (×107) TCID50/gram of tissue.

Based on the above, it was found that rhCC10 protein reduces the titer of virus in respiratory infections. This indicates the possibility of using rhCC10 protein to treat, cure and/or prophylaxis of influenza infection. In accordance with embodiments of the present invention provide based on rhCC10 protein therapy with intranasal and intravenous or combination thereof, effective to treat, cure or prevent influenza infection.

Example 4

The suppression of virus replication at the cellular level with the use of SS

Cells Ner ADS, Manassas, VA) were used for propagation of the virus RSV, strain A-2 (Advanced Biotechnologies, Inc., Columbia, MD), and obtaining a working viral drugs. Cells were sown in the amount of 50000 cells per cell in tablets of 48 cells and were grown in minimal medium eagle (MFM) with 10% serum embryo calves (FBS) to approximately 80% confluence. Cells pre-treated with protein S in 0.5 ml MEM for 4 hours. Then the medium was replaced by fresh one and the cells were infected with virus RSV in a dose of 1×106TCID50a Cup for cell cultures d�NetRom 100 mm for 1 hour. Neadsorbirovanne virus was removed by washing and added to 0.5 ml of medium MEM with 2% FBS, 4 mm L-glutamine and rhCC10 protein. Supernatant was collected on the fourth day after infection and virus was titrated. Fig.3 shows that the protein S at a concentration of 1 mg/ml almost completely eliminates the production of virus RSV, and at doses of 100 and 300 µg/ml was observed approximately 3-fold reduction.

Protein S also inhibited viral replication in the cells, if it was administered after 1, 24 and 48 hours after infection. Fig.4 shows that rhCC10 protein effective in suppressing the titer of the virus when administered not only before infection, but also after infection. This is the first direct evidence of antiviral activity of protein S at the cellular level and illustrates the potential benefits rhCC10 protein as a means of antiviral therapy in the treatment after infection.

Example 5

The mechanism of the antiviral action of protein S

The phenotype of epithelial cells of the respiratory tract in mice with a knockout for S shows that in the absence of protein S distribution of intracellular organelles abnormally that there was an anomalous way starannie membrane structure and violation of the secretion of other proteins produced by the cell. The authors suggest that this phenotype indicates that the protein S plays an active role in the transport of secretory vesicles from �of Parata Golgi to the plasma membrane of cells. Protein S also modulates the capture and processing of antigens to antigen-presenting cells. The authors suggest that these observations indicate that protein S is an important factor in the transport of materials both outside and inside many cell types. From this the authors conclude that protein S suppresses viral replication, preventing the transport of the virus into the cell. Since all viruses use cellular processes transport for introduction into the cell and replication, it should be expected that protein S can suppress the replication of all viruses. You can also expect that other secretogranin having a structure similar to a squirrel SS, also inhibit viral replication at the cellular level. Similarly, we can expect that obtained from protein S and other secretagogues, modulating cellular processes transport peptides are also able to suppress viral replication.

Links to related application

The present application is an international PCT application claiming priority to the provisional application U.S. No. 61/252028, filed October 15, 2009, the contents of which are fully incorporated in this application by reference.

1. The use of rhCC10 protein for the preparation of medicament for therapeutic or preventive effects on influenza virus.

2. The use according to claim 1, wherein �decreases the titer of virus in the tissue of the patient.

3. The use according to claim 1, characterized in that the influenza virus is an influenza virus type A.

4. The use according to claim 1, characterized in that the influenza virus is a strain of H1N1 influenza virus.

5. The use according to claim 2, characterized in that the tissue is a lung tissue.

6. The use according to claim 1, characterized in that the person or animal is inhibited viral replication at the cellular level.

7. The use according to claim 1, characterized in that suppressed viral replication in the infected cells at the cellular level.

8. The use according to any one of claims. 1-7, characterized in that rhCC10 protein affects influenza infections, cure or prevent influenza infection.

9. The use according to any one of claims. 1-7, characterized in that the protein rhCC10 is administered intranasal route.

10. The use according to any one of claims. 1-7, characterized in that the protein rhCC10 is administered intravenously by.

11. The use according to any one of claims. 1-7, characterized in that the protein rhCC10 is administered with the combination of intranasal and intravenous routes.



 

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21 cl, 2 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: invention relates to organic chemistry and particularly to aminoalkyl esters of 5-methoxyindole-3-carboxylic acid and pharmacologically acceptable salts thereof of general formula (I), where R1 is cyclohexyl, C1-3alkyl; R2 is phenylthio, phenyloxy, wherein the phenyl group can have 1-2 halogen substitutes or a C1-4alkoxy group, or R2 is a 5-6-member heterocycloalkyl containing 1-2 heteroatoms selected from nitrogen and oxygen; n equals 1, 2, 3, 4; each R is independently selected from C1-4alkyl; except compounds indicated in the claim. The invention also relates to a method of producing a compound of formula (I) and use thereof.

EFFECT: obtaining novel 5-methoxyindole-3-carboxylic acid derivatives, having antiviral activity.

4 cl, 2 tbl, 12 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed invention relates to such compositions and pharmaceutical compositions, which include poxviruses, and namely to those, which include extracellular enveloped viruses. Claimed invention also relates to such method, which is intended for production of poxviruses, as well as poxviruses, obtained in accordance with claimed invention. In addition, claimed invention also relates to application of claimed poxviruses and said composition for medication preparation.

EFFECT: obtaining pharmaceutical compositions, which include poxviruses.

11 cl, 3 dwg

FIELD: biotechnologies.

SUBSTANCE: method of obtaining of a complex of antimicrobic peptides of an insect includes infecting of adipose body of an insect at a larval instar with Micrococcus luteus A270 and Escherichia coli D31 bacteria with the subsequent extraction of adipose body of an insect at a larval instar. The adipose body of an insect is placed into a nutrient medium containing water solution of sugars, inorganic salts and the antibiotic meropenem in pre-set ratio and incubated during a day with the subsequent elution of the complex of antimicrobic peptides of an insect from cultural liquid by the method of reverse-phase chromatography on the column Vydac C18 at the linear gradient of acetonitrile from 0% up to 50%.

EFFECT: invention allows to simplify a method of obtaining antimicrobic peptides.

5 dwg, 4 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to biotechnology, namely to novel IL-17-inhibiting polypeptides, corresponding to fused proteins, to compositions and their application for medicinal purposes. Polypeptide contains amino acid sequence, which is selected from group, consisting of GVTLFVALYD YKAFWPGDLS FHKGEKFQIL RTSDGDWWEA RSLTTGETGY IPSNYVAPVD SIQ (SEQ ID NO: 39), GVTLFVALYD YKAFWPGDIS FHKGEKFQIL RTSDGEWWVA RSLTTGEEGY IPSNYVAPVD SIQ (SEQ ID NO: 57) or GVTLFVALYD YKAFWPGDIS FHKGEKFQIL RTSDGEWWIA RSLTTGEEGY IPSNYVAPVD SIQ (SEQ ID NO: 107); amino acid sequence, which has, at least, 80%, preferably, at least, 90%, more preferably, at least, 95% identity of amino acid sequence with SEQ ID NO: 39, SEQ ID NO:57 or SEQ ID NO: 107; fragment or functional derivative of SEQ ID NO: 39, SEQ ID NO: 57 or SEQ ID NO: 107, obtained due to substitution, addition and/or removal of not more than 5 amino acids.

EFFECT: invention makes it possible to bind IL-17 with high specificity and affinity.

33 cl, 17 dwg, 3 tbl, 12 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biotechnology and represents a cell-penetrating peptide for enhancing the passage of a hydrophilic physiologically active substance through a layer of mucosal epitheliocytes, as well as a pharmaceutical composition containing the above peptide. The cell-penetrating peptide represents a peptide with an amino acid sequence presented by SEQ ID NO: 1, as well as a peptide with an amino acid sequence presented by SEQ ID NO: 1, and having various modification; the peptide is able for penetrating through the cell membrane.

EFFECT: invention enables enhancing the passage of the hydrophilic physiologically active substance through the layer of mucosal epitheliocytes and making the hydrophilic physiologically active substance flow into the general circulation.

8 cl, 16 dwg, 1 tbl, 12 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to a pharmaceutical composition containing Ac-Arg-cyclo(Cys-D-Ala-His-D-Phe-Arg-Trp-Cys)-NH2 pamoate, which is a melanocortin receptor ligand subtype 4 (MC4-R) and in which after subcutaneous or intramuscular administration into an individual, the peptide forms a depot with physiologically value pH, which dissolves slowly and is released into body fluid and blood flow. The present invention also contains polyethylene glycol (PEG) at an average molecular weight from approximately 200 to approximately 400.

EFFECT: present invention possesses high effectiveness associated with a smooth release profile.

24 cl, 12 dwg, 6 tbl, 1 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to gastroenterology, and can be used for the treatment of chronic constipation and functional anorexia. For this purpose, as medicinal nutrition used is a milk-vitamin mixture with the following composition (g per 100 g of the product): Proteins 24-26, Fats 27-29, Carbohydrates 33-34, minerals (mg per 100 g of the product), calcium 940-970, phosphorus 780-820, sodium 230-270, potassium 1370-1550, chloride 1270-1350, magnesium 100-125, iron 9.5-10.7, zinc 2.7-3.5, iodine 145-173, copper 76-87, manganese 45-52, vitamins (mcg per 100 g of the product) D3 7.6-8.2, E 6.2-6.8, C 42-46, B1 960-990, B2 1150-1250, Niacin11-15, B6 1370-1440, Folic acid 125-150, Pantothenic acid 2250-2370, B12 1.5-1.9, Biotin 25-31, Choline 40-45.

EFFECT: invention provides an increase of treatment efficiency.

3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: object of the invention is a method and a pharmaceutical composition in the form of a water-alcohol solution of ethanol 30-60° and water 40-70%, wherein at least one hypoglycaemic active substance is stably and completely dissolved, to be used by administering through the oral mucosa as a therapeutic agent in accurate treatment of postprandial hyperglycemia accompanying type II diabetes mellitus in a human or animal; wherein the water-alcohol solution in the composition has a volume of less than 2 ml, wherein an amount of 250mg or less of the above active substance is stably and completely dissolved, while the hypoglycaemic active substance is specified in lipophilic or amphiphilic active substances, such as gliclazide, glinides, incretins and glyphins. The invention also refers to a method for preparing this dosage form.

EFFECT: preparing the therapeutic agent for treating postprandial hyperglycemia accompanying type II diabetes mellitus.

9 cl, 20 ex

FIELD: medicine.

SUBSTANCE: group of inventions refers to medicine and can be used for producing a controlled-release therapeutic agent containing a modified biologically active agent encapsulated into a polymer, wherein the above modified biologically active agent represents a peptide having an amino acid sequence specified in a group consisting of SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:9. What is also presented is providing the more effective charge of the biological agent, varying an erosion release rate of the biologically active agent, as well as varying the initial diffuse release of the biologically active agent in a polymer-based delivery system.

EFFECT: group of inventions enables changing release kinetics and/or parameters of the drug load of a peptide or a protein encapsulated in the polymer-based delivery systems through the direct variation of an isoelectric point and/or total charge of this peptide without changing the polymer charge.

13 cl, 10 dwg, 1 tbl, 2 ex

FIELD: agriculture.

SUBSTANCE: invention relates to agriculture, in particular to animal husbandry. The method of increasing productivity of cattle comprises adding to the animal diet of needle energy supplement at the rate of 250 g per head per day.

EFFECT: use of invention enables to increase milk production of cows.

2 tbl

FIELD: biotechnologies.

SUBSTANCE: polypeptide has a sequence represented by the formula below X1 Va1 X2 Ile Phe Thr Tyr Gly X3 Leu Gln Arg X4 Gln Glu Lys Glu Arg X5 Lys Pro Gln (formula I). Links between amino acids, apart from X1-Val link, are amide links. The X1-Val link is an amide link or a link represented by the formula below 2

X1 is an aromatic amino acid or a heteroaromatic amino acid, X2 represents proline or sarcosine, X3 is glutaminic acid or asparaginic acid, X4 is methionine or leucine, X5 is asparagine or proline. The proposed invention may be used in medicine for treatment of a disease related to disturbed function of gastrointestinal tract.

EFFECT: invention makes it possible to efficiently treat diseases related to disturbed function of gastrointestinal tract.

7 cl, 5 dwg, 4 tbl, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to pharmaceutics and medicine, and concerns protein and peptide stabilisation formulation which contains a hydrophilic polymer, a mixture of polyalcohol and sugar, wherein a weight ratio of polyalcohol to sugar makes from 2:1 to 5:1 (wt %), a detergent, and wherein the formulation is free from stabilising proteins. A composition and a kit for treating a disease or a composition caused by hyperactive cholinergic innervation of muscles or endocrine glands in a patient, and contain the above formulation and peptide, protein or a mixture thereof.

EFFECT: group of inventions provides the better protein stability in the absence of stabilising proteins.

13 cl, 4 ex

FIELD: medicine, purulent surgery.

SUBSTANCE: one should perform generally accepted medicinal therapy, moreover, one should prescribe additional chemotrypsin during the first 1-2 d after operation to be locally injected twice-thrice during day-time period at concentration of 0.5-1.0 mg/ml of 10%-sodium chloride solution at exposure of 1.5-2 h at the quantity of one fourth up to one third against the purulent volume removed out of abscess cavity, then therapy course should be supplemented with bacteriophage which should be locally injected twice during day-time period at daily dosage being 200 ml, not more at exposure of 1.5-2 h at the quantity of one tenth up to one fifth against purulent volume removed out of abscess cavity. As for bacteriophage type, it should be matched in accordance to the results of bacteriological survey of abscess cavitary content. Therapy course lasts for 6-9 d.

EFFECT: higher efficiency of therapy.

2 ex

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